pp 1-10 | Cite as

Total Protein Content Determination of Microalgal Biomass by Elemental Nitrogen Analysis and a Dedicated Nitrogen-to-Protein Conversion Factor

  • L. M. L. Laurens
  • J. L. Olstad
  • D. W. Templeton
Protocol
Part of the Methods in Molecular Biology book series

Abstract

Accurately determining protein content is important in the valorization of algal biomass in food, feed, and fuel markets, where these values are used for component balance calculations. Conversion of elemental nitrogen to protein is a well-accepted and widely practiced method, but depends on developing an applicable nitrogen-to-protein conversion factor. The methodology reported here covers the quantitative assessment of the total nitrogen content of algal biomass and a description of the methodology that underpins the accurate de novo calculation of a dedicated nitrogen-to-protein conversion factor.

Keywords

Amino acids Nitrogen analysis Nitrogen-to-protein conversion factor Protein 

Notes

Acknowledgments

This work was supported by the U.S. Department of Energy under Contract No. DE-AC36-08GO28308 with the National Renewable Energy Laboratory. Funding provided by U.S. DOE Office of Energy Efficiency and Renewable Energy Bioenergy Technologies Office. The U.S. Government retains and the publisher, by accepting the article for publication, acknowledges that the U.S. Government retains a nonexclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this work, or allow others to do so, for U.S. Government purposes.

References

  1. 1.
    Becker EW (1994) Microalgae—biotechnology and microbiology. Cambridge University Press, CambridgeGoogle Scholar
  2. 2.
    Pignolet O, Jubeau S, Vaca-Garcia C, Michaud P (2013) Highly valuable microalgae: biochemical and topological aspects. J Ind Microbiol Biotechnol 40:781–796.  https://doi.org/10.1007/s10295-013-1281-7CrossRefGoogle Scholar
  3. 3.
    Lourenço S, Barbarino E (1998) Distribution of intracellular nitrogen in marine microalgae: basis for the calculation of specific nitrogen-to-protein conversion factors. J Phycol 811:798–811Google Scholar
  4. 4.
    Templeton DW, Laurens LML (2015) Nitrogen-to-protein conversion factors revisited for applications of microalgal biomass conversion to food, feed and fuel. Algal Res 11:359–367.  https://doi.org/10.1016/j.algal.2015.07.013CrossRefGoogle Scholar
  5. 5.
    Koru E (2012) Earth food Spirulina (Arthrospira): production and quality standarts. In: El-Samragy Y (ed) Food addit. InTech, Rijeka, pp 191–203Google Scholar
  6. 6.
    Garcia MDC, Lopez CVG, Fernandez FGA et al (2010) Protein measurements of microalgal and cyanobacterial biomass. Bioresour Technol 101:7587–7591.  https://doi.org/10.1016/J.Biortech.2010.04.077CrossRefGoogle Scholar
  7. 7.
    Servaites JC, Faeth JL, Sidhu SS (2012) A dye binding method for measurement of total protein in microalgae. Anal Biochem 421:75–80.  https://doi.org/10.1016/j.ab.2011.10.047Google Scholar
  8. 8.
    Slocombe SP, Ross M, Thomas N et al (2013) A rapid and general method for measurement of protein in micro-algal biomass. Bioresour Technol 129:51–57.  https://doi.org/10.1016/j.biortech.2012.10.163CrossRefGoogle Scholar
  9. 9.
    Redinbaugh MG, Turley RB (1986) Adaptation of the bicinchoninic acid protein assay for use with microtiter plates and sucrose gradient fractions. Anal Biochem 153:267–271Google Scholar
  10. 10.
    Laurens LML, Van Wychen S, McAllister JP et al (2014) Strain, biochemistry, and cultivation-dependent measurement variability of algal biomass composition. Anal Biochem 452:86–95.  https://doi.org/10.1016/j.ab.2014.02.009CrossRefGoogle Scholar
  11. 11.
    AOAC (1997) Amino acids in feeds. Official Method 994.12Google Scholar
  12. 12.
    Mosse J (1990) Nitrogen-to-protein conversion factor for ten cereals and six legumes or oilseeds. A reappraisal of its definition and determination. Variation according to species and to seed protein content. J Agric Food Chem 38:18–24.  https://doi.org/10.1021/jf00091a004CrossRefGoogle Scholar
  13. 13.
    Lourenço SO, Barbarino E, Marquez UML, Aidar E (1998) Distribution of intracellular nitrogen in marine microalgae: Basis for the calculation of specific nitrogen-to-protein conversion factors. J Phycol 34:798–811Google Scholar
  14. 14.
    Lourenço SO, Barbarino E, Lavín PL et al (2004) Distribution of intracellular nitrogen in marine microalgae: calculation of new nitrogen-to-protein conversion factors. Eur J Phycol 39:17–32.  https://doi.org/10.1080/0967026032000157156CrossRefGoogle Scholar
  15. 15.
    Lourenco SO, Barbarino E, De-Paula JC et al (2002) Amino acid composition, protein content and calculation of nitrogen-to-protein conversion factors for 19 tropical seaweeds. Phycol Res 50:233–241.  https://doi.org/10.1046/j.1440-1835.2002.00278.xCrossRefGoogle Scholar
  16. 16.
    AOAC (1990) Protein (Crude) in animal feed: combustion method. Official Method 990.03Google Scholar
  17. 17.
    Ball MF, Meloni CR, Diaz MJ et al (1966) Nitrogen measurement in biological fluids and bone by an automatic Dumas technique. Anal Biochem 16:343–348Google Scholar
  18. 18.
    Kjeldahl J (1883) Neue Methode zur Bestimmung des Stickstoffs in organischen Körpern. Zeitschrift fur Anal Chemie 22:366–383Google Scholar

Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  • L. M. L. Laurens
    • 1
  • J. L. Olstad
    • 1
  • D. W. Templeton
    • 1
  1. 1.National Renewable Energy LaboratoryNational Bioenergy CenterGoldenUSA

Personalised recommendations